Investigation of the role of miRNA targeting NFAT as a biomarker of Myocardial Infarction

Abstract

According to the World Heart Report 2023, 85% of the total cardiovascular diseases associated deaths are caused by coronary artery disease and stroke that is 9.1 million deaths approximately. Coronary artery disease is a complex disorder that involves occlusion of coronary artery, inflammation, and apoptosis of the affected part of the heart. Moreover, there is dysregulation of the microRNAs and their associated target gene signaling pathways. These miRNAs may serve as the diagnostic biomarker for coronary artery disease. This study was aimed to investigate the use of miR-1-3p and miR-98-5p as the potential biomarker for coronary artery disease diagnosis. This study also tried to put forward a novel fluorometric strategy for miRNA quantification. We found an elevated expression of miR-1-3p and decreased expression of miR-98-5p in whole blood samples of coronary artery disease patients. Fluorescence intensity-based quantification of miR-1-3p also showed a remarkably increased quantity of this miRNA in CAD patients. To validate the expression of these miRNAs and their target genes, animal model studies were also conducted. Oxidants and antioxidants biochemical assays confirmed oxidative stress in ISO induced rats. Triglycerides and cholesterol levels were found to be high among the diseased group. Liver markers including activities of AST and ALT were also increased. In the blood and tissue samples of diseased model, Et-1, Bcl-2, and BAX mRNA levels were found to be significantly elevated while NFATc3 mRNA levels were increased in the tissue samples only. To confirm ET-1 as target of miR-98-5p, western blotting was performed after dosing miRNA-98-5p mimics. Upon administration of miRNA-98-5p mimic, Et-1, Drp-1, NFATc3, and BAX protein expression levels were decreased in the diseased group. Furthermore, the histological analysis by H&E staining depicted altered cellular morphology with an elevated percentage of abnormal cells in the diseased group. In conclusion, miR-1-3p and miR-98-5p may serve as potential non-invasive diagnostic biomarkers for coronary artery disease because of their regulatory roles in critical apoptotic and hypertrophic pathways.